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. 2021 Jun 2;8:674747. doi: 10.3389/fmolb.2021.674747

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Copyright © 2021 Hoyt.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Quantitative assessment of equivalence to chromogenic IHC staining. Staining parameters for each antibody are first optimized using single stain, chromogenic IHC on tonsil sections. Next, each primary antibody was paired to a select TSA fluorophore and a single-stain, monoplex IF (monoIF) was performed. Lastly, all of the monoplex conditions are combined into a multiplex IF (mIF) panel. (A) Representative image showing serial sections with each marker for IHC, monoIF, and mIF staining. (B) Quantitative assessment of each image demonstrating the equivalence of staining for each marker across the three staining parameters, wherein monoIF and mIF are consistent with DAB.